visualization-quantification-cell-free-layer-arterioles-rat-cremaster

Visualization and Quantification of the Cell-free Layer in Arterioles of the Rat Cremaster Muscle

The cell-free layer is defined as the parietal plasma layer in the microvessel flow, which is devoid of red blood cells. The measurement of the in vivo ...

National University of Singapore

Computational modeling of Red Blood Cell (RBC) flow contributes to the fundamental understanding of microhemodynamics and microcirculation. In order to construct theoretical RBC models, experimental studies on single RBC mechanics have presented a material description for RBC membranes based on their membrane shear, bending and area moduli. These properties have been directly employed in 3D continuum models of RBCs but practical flow analysis with 3D models have been limited by their computationally expensive nature. As such, various researchers have employed 2D models to efficiently and qualitatively study microvessel flows. Currently, the representation of RBC dynamics using 2D models is a limited methodology that breaks down at high shear rates due to excessive and unrealistic stretching.

Two-dimensional strain-hardening membrane model for large deformation behavior of multiple red blood cells in high shear conditions.

Despite the significant roles of the cell-free layer (CFL) in balancing nitric oxide (NO) and oxygen (O2) bioavailability in arteriolar tissue, many previous numerical approaches have relied on a one-dimensional (1-D) steady-state model for simplicity. However, these models are unable to demonstrate the influence of spatiotemporal variations in the CFL on the NO/O2 transport under dynamic flow conditions. Therefore, the present study proposes a new two-dimensional (2-D) transient model capable of predicting NO/O2 transport modulated by the spatiotemporal variations in the CFL width. Our model predicted that NO bioavailability was inversely related to the CFL width as expected. The enhancement of NO production by greater wall shear stress with a thinner CFL could dominate the diffusion barrier role of the CFL. In addition, NO/O2 availability along the vascular wall was inhomogeneous and highly regulated by dynamic changes of local CFL width variation. The spatial variations of CFL widths on opposite sides of the arteriole exhibited a significant inverse relation. This asymmetric formation of CFL resulted in a significantly imbalanced NO/O2 bioavailability on opposite sides of the arteriole. The novel integrative methodology presented here substantially highlighted the significance of spatiotemporal variations of the CFL in regulating the bioavailability of NO/O2, and provided further insight about the opposing effects of the CFL on arteriolar NO production.

Two-dimensional transient model for prediction of arteriolar NO/O2 modulation by spatiotemporal variations in cell-free layer width.

This study examined the effect of dextran-induced RBC aggregation on the venular flow in microvasculature. We utilized the laser speckle contrast imaging (LSCI) as a wide-field imaging technique to visualize the flow distribution in venules influenced by abnormally elevated levels of RBC aggregation at a network-scale level, which was unprecedented in previous studies. RBC aggregation in rats was induced by infusing Dextran 500. To elucidate the impact of RBC aggregation on microvascular perfusion, blood flow in the venular network of a rat cremaster muscle was analyzed with a stepwise reduction of the arterial pressure (100 → 30 mmHg). The LSCI analysis revealed a substantial decrease in the functional vascular density after the infusion of dextran. The relative decrease in flow velocity after dextran infusion was notably pronounced at low arterial pressures. Whole blood viscosity measurements implied that the reduction in venular flow with dextran infusion could be due to the elevation of medium viscosity in high shear conditions (> 45 s-1). In contrast, further augmentation to the flow reduction at low arterial pressures could be attributed to the formation of RBC aggregates (< 45 s-1). This study confirmed that RBC aggregation could play a dominant role in modulating microvascular perfusion, particularly in the venular networks.

Alteration of Blood Flow in a Venular Network by Infusion of Dextran 500: Evaluation with a Laser Speckle Contrast Imaging System.

This study examined the effect of red blood cell (RBC) aggregation on nitric oxide (NO) and oxygen (O2) distributions in the downstream vessels of arteriolar bifurcations. Particular attention was paid to the inherent formation of asymmetric cell-free layer (CFL) widths in the downstream vessels and its consequential impact on the NO/O2 bioavailability after the bifurcations. A microscopic image-based two-dimensional transient model was used to predict the NO/O2 distribution by utilizing the in vivo CFL width data obtained under non-, normal- and hyper-aggregating conditions at the pseudoshear rate of 15.6±2.0s(-1). In vivo experimental result showed that the asymmetry of CFL widths was enhanced by the elevation in RBC aggregation level. The model demonstrated that NO bioavailability was regulated by the dynamic fluctuation of the local CFL widths, which is corollary to its modulation of wall shear stress. Accordingly, the uneven distribution of NO/O2 was prominent at opposite sides of the arterioles up to six vessel-diameter (6D) away from the bifurcating point, and this was further enhanced by increasing the levels of RBC aggregation. Our findings suggested that RBC aggregation potentially augments both the formation of asymmetric CFL widths and its influence on the uneven distribution of NO/O2 in the downstream flow of an arteriolar bifurcation. The extended heterogeneity of NO/O2 downstream (2D-6D) also implied its potential propagation throughout the entire arteriolar microvasculature.

Erythrocyte aggregation may promote uneven spatial distribution of NO/O2 in the downstream vessel of arteriolar bifurcations.

Heterogeneous distribution of red blood cells (RBCs) in downstream vessels of arteriolar bifurcations can be promoted by an asymmetric formation of cell-free layer (CFL) in upstream vessels. Consequently, the CFL widths in subsequent downstream vessels become an important determinant for tissue oxygenation (O2) and vascular tone change by varying nitric oxide (NO) availability. To extend our previous understanding on the formation of CFL in arteriolar bifurcations, this study investigated the formation of CFL widths from 2 to 6 vessel-diameter (2D-6D) downstream of arteriolar bifurcations in the rat cremaster muscle (D = 51.5 ± 1.3 μm). As the CFL widths are highly influenced by RBC aggregation, the degree of aggregation was adjusted to simulate levels seen during physiological and pathological states. Our in vivo experimental results showed that the asymmetry of CFL widths persists along downstream vessels up to 6D from the bifurcating point. Moreover, elevated levels of RBC aggregation appeared to retard the recovery of CFL width symmetry. The required length of complete symmetry recovery was estimated to be greater than 11D under reduced flow conditions, which is relatively longer than interbifurcation distances of arterioles for vessel diameter of ∼50 μm. In addition, our numerical prediction showed that the persistent asymmetry of CFL widths could potentially result in a heterogeneous vasoactivity over the entire arteriolar network in such abnormal flow conditions.

Article	Year
Two-dimensional strain-hardening membrane model for large deformation behavior of multiple red blood cells in high shear conditions. Theoretical biology & medical modelling\| PubMed ID: 24885482	2014
Two-dimensional transient model for prediction of arteriolar NO/O2 modulation by spatiotemporal variations in cell-free layer width. Microvascular research\| PubMed ID: 25312045	2015
Alteration of Blood Flow in a Venular Network by Infusion of Dextran 500: Evaluation with a Laser Speckle Contrast Imaging System. PloS one\| PubMed ID: 26466371	2015
Erythrocyte aggregation may promote uneven spatial distribution of NO/O2 in the downstream vessel of arteriolar bifurcations. Journal of biomechanics\| PubMed ID: 26684432	2016
Symmetry recovery of cell-free layer after bifurcations of small arterioles in reduced flow conditions: effect of RBC aggregation. American journal of physiology. Heart and circulatory physiology\| PubMed ID: 27233764	2016

Yan Cheng Ng

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